| Objective(1)To explore the effect of Effective Components of Qi-toads Colon-specific Oral Tablets(ECOQCOT)on CT-26 tumor-bearing mice,and further research the effect of ECOQCOT on T-lymphocytes level in the peripheral blood of tumor-bearing mice,the expression of related apoptosis regulatory factors Bax,Bcl-2 and angiogenesis related factors VEGF,HIF-1α in transplanted tumor tissue,in order to explore the mechanism of ECOQCOT on colonic carcinoma.(2)To explore the effect of ECOQCOT combine with fluorouracil(5-FU)on CT-26 tumor-bearing mice during its process of arising and developing.To understand the optimal timing to intervene and the synergy and attenuation effect of ECOQCOT when combine with 5-FU through researching the effect of ECOQCOT combine with 5-FU on T-lymphocytes level in the peripheral blood of tumor-bearing mice,the expression of related apoptosis regulatory factors Bax,Bcl-2 and angiogenesis related factors VEGF,HIF-1α in transplanted tumor tissue.To provides the theory basis for clinical prevention and rehabilitation of colonic carcinoma.Methods(1)The mechanism of ECOQCOT on CT-26 transplanted colonic tumor-bearing mice:Cell suspension subcutaneous inoculation method is adopted to establish the CT-26 transplanted colonic tumor model,and then the mice were divided into normal group,model group,5-FU group,high-dose ECOQCOT group and low-dose ECOQCOT group.(2)Effect of ECOQCOT combine with 5-FU on CT-26 transplanted colonic tumor-bearing mice:Cell suspension subcutaneous inoculation method is adopted to establish the CT-26 transplanted colonic tumor model,then the mice were divided into model group,early intervention group,synchronous intervention group,delayed intervention group,5-FU group and ECOQCOT group.The two parts above both observing the general situation of mice during the period of drug delivery;dissecting the mice after the last drug delivery,getting the tumor,spleen and thymus,to calculate tumor inhibitory rate and the spleen and thymus index;using HE staining method to observe the pathologic morphology of transplantation tumor tissue in each group,detecting the level of CD3+,CD4+,CD8+T-lymphocytes and CD4+/CD8+in the peripheral blood by flow cytometry,adopting immunohistochemical staining to detect the expressions of Bax,Bcl-2,VEGF,HIF-1α proteins of transplantation tumor tissue.Results(1)The mechanism of ECOQCOT on CT-26 transplanted colonic tumor-bearing mice.①Antitumor effects:The tumor inhibition rate of high-dose ECOQCOT group and low-dose ECOQCOT group was 33.90%and 26.72%.Compared with model group,tumor weight of high-dose ECOQCOT group and low-dose ECOQCOT group were significantly reduced(P<0.01).②The effect of ECOQCOT on immune organ index in mice:Compared with model group,spleen index in ECOQCOT groups were increased.The thymus index in high-dose ECOQCOT and low-dose ECOQCOT group were higher than that in model group(P<0.01,P<0.05).③The expression of T-lymphocytes in peripheral blood in mice:Compared with model group,cell proportion of CD3+,CD4+,CD8+and CD4+/CD8+ were significantly increased in high-dose ECOQCOT group(P<0.01);and cell proportion of CD3+and CD4+/CD8+were significantly increased in low-dose ECOQCOT group(P<0.05).④The expression of related apoptosis regulatory factors in transplanted tumor tissue:The IOD of Bax in high-dose ECOQCOT group and low-dose ECOQCOT group were increased,compared with model group,the IOD of Bax in high-dose ECOQCOT group was significantly increased(P<0.05);the IOD of Bcl-2 in high-dose ECOQCOT group and low-dose ECOQCOT group were reduced,compared with model group,the IOD of Bcl-2 in high-dose ECOQCOT group was significantly reduced(P<0.01).⑤The expression of angiogenesis related factors in transplanted tumor tissue:The IOD of VEGF and HIF-1α in high-dose ECOQCOT group and low-dose ECOQCOT group were reduced.Compared with model group,the expression of VEGF,HIF-1α protein in high-dose ECOQCOT group were reduced,difference were statistically significant(P<0.01,P<0.05).(2)Effect of ECOQCOT combine with 5-FU on CT-26 transplanted colonic tumor-bearing mice.①Antitumor effects:Compared with model group,tumor weight of early intervention group,synchronous intervention group,delayed intervention group,5-FU group and ECOQCOT group were significantly reduced(P<0.01,P<0.01,P<0.01,P<0.01,P<0.05);tumor weight of early intervention group was significantly lower than that of 5-FU group and ECOQCOT group,difference were statistically significant(P<0.5,P<0.01).②The immune organ index in mice:There were no significantly difference between groups.③ The expression of T-lymphocytes in peripheral blood in mice:Compared with model group,cell proportion of CD3+,CD4+,CD8+were increased in early intervention group,synchronous intervention group,delayed intervention group,5-FU group and ECOQCOT group,difference were statistically significant(P<0.01);and CD4+/CD8+in early intervention group and synchronous intervention group were significantly higher than that in model group(P<0.01).Compared with 5-FU group,cell proportion of CD3+,CD4+in early intervention group,synchronous intervention group and delayed intervention group were significantly increased(P<0.01).Compared with ECOQCOT group,cell proportion of CD3+,CD4+and CD4+/CD8+were significantly increased in early intervention group(P<0.05,P<0.01,P<0.01),cell proportion of CD4+and CD4+/CD8+in synchronous intervention group were significantly higher than that in ECOQCOT group(P<0.05).④The expression of related apoptosis regulatory factors in transplanted tumor tissue:Compared with model group,the IOD of Bax in early intervention group,synchronous intervention group were increased,difference were statistically significant(P<0.01,P<0.05).Compared with model group,the IOD of Bcl-2 in early intervention group,synchronous intervention group,delayed intervention group and 5-FU group were reduced,difference were statistically significant(P<0.01,P<0.01,P<0.05,P<0.05).⑤The expression of angiogenesis related factors in transplanted tumor tissue:Compared with model group,expression of VEGF protein in early intervention group and synchronous intervention group were reduced,difference were statistically significant(P<0.01,P<0.05).Compared with model group,the IOD of HIF-1α in each treatment group were reduced,the IOD of HIF-1α in early intervention group,synchronous intervention group and delayed intervention group were lower than that in 5-FU group and ECOQCOT group.Conclusion(1)The mechanism of ECOQCOT on CT-26 transplanted colonic tumor-bearing mice:ECOQCOT has inhibitive effects on CT-26 transplanted colonic tumor-bearing mice,it can increase the expression of T-lymphocytes in peripheral blood of mice;increase the expression of Bax protein,decrease the expression of Bcl-2,VEGF,HIF-lα proteins.Exert an anti-tumor effect through adjusting the immune fuction of transplanted tumor-bearing mice,inducing tumor cells apoptosis and inhibiting angiogenesis.(2)Effect of ECOQCOT combine with 5-FU on CT-26 transplanted colonic tumor-bearing mice:The efficacy of ECOQCOT combine with 5-FU groups were better than 5-FU group and ECOQCOT group.The combination regimen groups had synergetic effects,they can reduce the adverse reaction of 5-FU,early intervention group had the optimal effect.These actions may be related to its action of enhancing the immune fuction of transplanted tumor-bearing mice,inducing tumor cells apoptosis and inhibiting angiogenesis. |
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